Flag-tag Rapid Assay (Colloidal Gold), 10 T

Product Information

Product Description/Introduction

The Flag-tag Rapid Assay(colloidal gold) is a 10-minute dipstick lateral-flow assay to detect Flag-tagged proteins in cell culture and lysates during protein expression or in purified protein preparations. The assay is performed by simply applying the lateral flow strip into the sample of properly diluted tissue culture supernatant or lysate and visualizing via a loss of red bands in the test line section.

The product mainly consists of a sample pad, binding pad, chromatography membrane, absorbent paper, and backing plate. In a negative result where no Flag-tagged protein is present or concentration of Flag-tagged protein is below detectable levels, the gold-conjugated capture antibodies will bind to the Flag-tagged protein antigen embedded on the test lines and form a visible red line. In both positive and negative tests, the goldconjugated anti-Flag capture antibodies will bind to the control antibodies at the control line. In a positive test where the sample contains Flag-tagged proteins, the gold-conjugated antibodies embedded in the sample pad will bind to the available Flag-tag proteins in the sample and therefore not bind to the Flag-tagged protein antigens immobilized on the test lines. As the concentration increases, the color of test line will decrease until disappear. The concentration of the Flag-tagged proteins is inversely related to the color of test line appearing on the strip.

Storage and Shipping Conditions

Ship and store at room temperature, valid for 12 months.

Product Components

Assay Procedures/Protocols

1. Allow the package to warm to room temperature prior to removing the strips from the bag.

2. For each strip, add 80-100 μL of culture supernatant, lysate or purified protein preparations to a test tube.

3. Wait 8-10 minutes for the color bands to appear, it is invalid after 15 minutes.

Results Judgement

1. Negative result: Both the control line and the test line are coloured, with the test line showing the darkest colour.

2. Positive result: The control line is coloured, the test line is weakly coloured or not coloured.

3. Invalid result: When the control line is not coloured, the test should be repeated whether the test line is coloured.

Note: When the sample does not contain Flag-tag protein, the color of the test line is darkest, and gradually decreases as the amount of Flag-tag protein in the sample increases, until the colour disappears.

Results Display

Dilute the Flag-tagged protein with PBS to 10 μg/mL, 20 μg/mL and 50 μg/mL respectively, and then add 100 μL of dilution to the wells and leave it for 8 min.

Results of Flag-tag Rapid Assay(colloidal gold) for 8 min

Sensitivity

The lower limit of detection is 20 μg/mL, when the Flag-tag protein content in a sample is below 20 μg/mL, it is not easily distinguish with the naked eye.

Note

1. If the colour of the sample causes a high background, the sample can be diluted with PBS or culture medium before chromatography.

2. For the accuracy of the results, a negative control and positive control should be set up, and it is recommended that the control sample is the same matrix as the sample to be tested.

3. This test card is not recommended for freezing.

4. Once opened please use quickly.

For Research Use Only!

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